Kaeffer B, van Kooten P, Ederveen J, van Eden W, Horzinek M C
Department of Infectious Diseases and Immunology, Veterinary Faculty, State University of Utrecht, the Netherlands.
Am J Vet Res. 1989 Jul;50(7):1131-7.
Seven hybridomas that secreted monoclonal antibodies (MAB) against the peplomer protein and one that secreted MAB against the nucleocapsid protein of Berne virus (proposed family Toroviridae) were isolated. All MAB directed against the peplomer protein neutralized virus infectivity and, with the exception of MAB 6A7, inhibited each other's binding in competition assays. Neutralization of Berne virus infectivity was potentiated when some MAB were used in pairs. The antibodies have been used to localize toroviral proteins in infected cells; use of antipeplomer MAB 6B10 yielded a diffuse intracytoplasmic immunofluorescence, whereas the antinucleocapsid MAB 1F1 detected antigen in the intra- and perinuclear compartments. By use of radioimmune precipitation, protein A of Staphylococcus aureus was found to bind directly to the nucleocapsid polypeptide, without the requirement for specific antibody. Using fluorescein isothiocyanate-conjugated protein A, the intranuclear accumulation of the nucleoprotein of Berne virus was confirmed by results of immunofluorescence.
分离出7株分泌针对伯尔尼病毒(提议的环曲病毒科)纤突蛋白的单克隆抗体(MAB)的杂交瘤和1株分泌针对其核衣壳蛋白的MAB的杂交瘤。所有针对纤突蛋白的MAB均中和病毒感染性,并且除MAB 6A7外,在竞争试验中相互抑制结合。当一些MAB成对使用时,伯尔尼病毒感染性的中和作用增强。这些抗体已用于在感染细胞中定位环曲病毒蛋白;使用抗纤突MAB 6B10产生弥漫性胞浆内免疫荧光,而抗核衣壳MAB 1F1在核内和核周区室中检测到抗原。通过放射免疫沉淀法,发现金黄色葡萄球菌蛋白A直接结合到核衣壳多肽上,而无需特异性抗体。使用异硫氰酸荧光素偶联的蛋白A,通过免疫荧光结果证实了伯尔尼病毒核蛋白在核内的积累。